Project: MUSST - “Multi-step Streptamer systems for clinical stem cell research”
Stem cells are widely considered to have a huge potential for future clinical application, e.g. to treat incurable diseases like Parkinson´s or diabetes (Daley, GQ2012). During the last decade, it became evident that techniques for isolation of different stem cell populations are crucial for the translation of stem cells from bench to bedside. Clinical-grade magnetic bead-based isolation of cell populations bearing specific surface markers has become the most widely used option to purify stem cells from tissues/body fluids, with the Miltenyi CliniMACS system dominating the market world-wide. Currently, dozens of early stage clinical trials are performed, using stem cell preparations defined by the expression of single surface markers. Single surface markers often are not adequate to purify/define stem cell populations and that more surface markers have to be considered (Baum, DM et al 1992; Majeti R et al 2007; Vanheusden, K et al 2007; Notta, F et al 2011). Autologous hematopoietic stem cells (HSC) from mobilized peripheral blood (MPB) are frequently used to aid in hematopoietic reconstitution after high-dose chemotherapy. In many cases, contaminating cancer cells are detectable in the autologous MPB graft, making these patients prone to relapse. Transplantation of highly purified hematopoietic stem cells was demonstrated to have the potential to overcome this problem (Michallet, M et al.2000; Vose JM et al 2001), but as HSC purification is a complex, laborious and expensive task, these results did not have an impact on current clinical practice. For scientific purposes, multi-parameter FACS sorting is used to enable strong enrichment of e.g. hematopoietic stem cells. For the clinical application, this technique still requires investments of several 100000 € for FACS sorting equipment and poses various problems such as fluidics system / flow cell clean-up and aerosol formation (McIntyre CA et al.; 2010). In addition, as the antibodies reCO bound to the cells after sorting, these reagents are no ancillary materials, but have to fulfill GMP standards, which strongly limits the available antibody repertoire for clinical grade cell sorting. Magnetic bead based sorting, on the other hand, is not so cost intense, but does not allow multiple positive selection steps so far. This major limitation will be addressed by the MUSST project by developing a CD34/CD90 double positive selection kit based on IBA´s proprietary Streptamer technology. To enable clinical-grade cell separation, pre-existing know how on microfluidic-based cell separation (MABAZELL-Project, IBA and iba) and stem cell cultivation in closed-system pipe-based bioreactors (HYPERLAB Project, iba and MUW) and magnetofection (IBA, MATra System) will be developed to allow cell separation, transfection and cultivation in a closed system, which will be relevant for future clinical applications. P MUW will provide umbilical cord blood samples and know-how on hematopoetic stem cell isolation, characterization and cultivation together with flow cytometry and cell sorting expertise. The hematopoietic system was chosen, as it clearly represents the biggest market for stem cell separation reagents and is the best characterized system with clinical relevance. _x000D_Genetic modification of hematopoietic stem cells was the paradigm of gene therapy. It is also considered an option to optimize engraftment of transplanted hematopoietic stem cells. Transfer of drug resistance genes to hematopoietic cells before transplantation is envisaged as a new possibility to limit myelo-suppression after chemotherapy and enable higher drug concentrations for better anti-tumor treatment (Bertino JR 2008; Létourneau S et al 1996; Moritz, T and Williams, DA 2002). All these manipulations require ex-vivo manipulation of stem cells and are tightly regulated. A closed system, which can be COtained during HSC isolation and genetic manipulation, would be a major advance for the ultimate clinical use of genetically modified stem cells. The MUSST project will adapt existing know-how for microfluidics (iba) and magnetofection (IBA) to the specific purpose of hematopoietic stem cell transfection with RNA/DNA for possible therapeutic purposes. If the multi-step Streptamer platform is accepted by the market, this development could also be commercialized in the form of a cell isolation/transfection service or as a second application of the microfluidics-based closed system. The microfluidics-based closed system for cell separation, which will be developed during the MUSST project will also be of high commercial interest. _x000D_The multi-step Streptamer platform will not be restricted to the CD34/CD90 double positive selection kit, but can be adapted to allow multi-step positive selection of different surface antigens if required by the market/customers. _x000D_
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Acronym
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MUSST
(Reference Number: 8099)
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Duration
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01/01/2014 - 31/12/2016
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Project Topic
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Project aim is to develop a reversible multi-step cell separation technology for clinical stem cell research using the innovative Streptamer technology. The Streptamer technology uses magnetic nano- or microbeads which can be retained after each single purification step based on its reversibility._x000D_
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Network
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Eurostars
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Call
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Eurostars Cut-Off 10
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Project partner
